Molecular detection of Mycoplasma synoviae and avian reovirus infection in arthritis and tenosynovitis lesions of broiler and breeder chickens in Santa Catarina State, Brazil (original) (raw)
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Mycoplasma synoviae and avian reovirus (ARV) are associated with several disease syndromes in poultry and cause notable global economic losses in the poultry industry. Rapid and efficient diagnostics for these avian pathogens are important not only for disease control but also for prevention of clinical disease progression. However, current diagnostic methods used for surveillance of these diseases in poultry flocks are laborious and time-consuming, and they have low sensitivity. The multiplex PCR (mPCR) developed in this study has been proven to be both sensitive and specific for simultaneous M. synoviae and ARV detection and identification in clinical samples. To evaluate the mPCR assay, the diagnostic test was applied to different clinical samples from natural and experimental M. synoviae and ARV-infected poultry. Results were compared with serologic, single PCR, and immunofluorescence analyses. Tibiotarsal articulation could be the best target for simultaneous detection of M. synoviae and ARV infection. The detection limit by visualization of mPCR-amplified products was 100 pg for both pathogens. Overall, the mPCR developed and standardized in this research is a useful tool for diagnosis and screening and for surveillance and control of M. synoviae and ARV infection in poultry flocks.
Veterinary World, 2019
Aim: Arthritis is one of the most economic problems facing poultry industry worldwide. The study was done to detect possible causes of arthritis in breeder chicken flock with emphasis on molecular identification of Mycoplasma synoviae (MS). Materials and Methods: This study was carried on chicken from broiler breeder flock of 57 weeks' age in Dakahlia, Egypt, suffered from arthritis with frequently 5-7% decrease in egg production, reduced fertility, and hatchability. Forty blood samples were randomly collected from individual birds in sterile tubes and used for serum separation. Serum samples were tested using serum plate agglutination (SPA) test against colored antigens for Mycoplasma gallisepticum (MG), MS, and Salmonella gallinarum-pullorum (SGP). On the other hand, 24 joint samples were collected. Of those 24 samples, 12 joint samples were subjected to bacteriological examination, while the other 12 were utilized for molecular diagnosis by polymerase chain reaction (PCR) for MS and avian reovirus (ARV). Results: SPA test results revealed the presence of antibodies against MG, MS, and SGP in tested sera in rates of 14/40 (35%), 35/40 (87.5%), and 9/40 (22.5%), respectively. Furthermore, 19 bacterial isolates were recognized from joint samples and identified as five Staphylococcus spp., nine Escherichia coli, three SGP, one Citrobacter, and one Proteus. The identified Staphylococcal isolates were three coagulase-positive staphylococci (two Staphylococcus aureus and one Staphylococcus hyicus) and two coagulase-negative staphylococci (one Staphylococcus epidermidis and one Staphylococcus lentus), while E. coli isolate serotypes were 1 O11, 2 O55, 3 O78, 1 O124, 1 O125, and 1 untyped. PCR proved that 12/12 (100%) samples were positive for MS variable lipoprotein hemagglutinin A (vlhA) gene, while ARV was not diagnosed in any of the examined samples. Four amplified vlhA gene of MS isolates (named MS-2018D1, MS-2018D2, MS-2018D3, and MS-2018D4) was successfully sequenced. Analysis of phylogenetic tree revealed the presence of 100% identity between each two sequenced isolates (isolates MS-2018D1 and MS-2018D4 and also isolates 2018D2 and MS-2018D3). However, the nucleotide similarity between four isolates was 88.6%. On the other hand, our field isolates MS-2018D1, MS-2018D4, MS-2018D2, and MS-2018D3 showed nucleotide identity with vaccine strain MS-H 98.4%, 98.4%, 88.1%, and 88.1%, respectively. Furthermore, the nucleotide similarities with field strains from Argentina ranged between 87.8% and 98.6%. Conclusion: Four field isolates of MS were identified in examined broiler breeder flock. A phylogenetic study of these isolates revealed the variation between isolated MS strains and vaccine strain. Therefore, further studies are required for evaluating the vaccine efficacy against the present field isolates of MS. In addition, application of MS immunization of breeder flocks is necessary for proper control of the disease.
Pathogenicity of newly emergent turkey arthritis reoviruses in chickens
Poultry Science, 2015
Turkey arthritis reoviruses (TARVs) were isolated recently from gastrocnemius and digital flexor tendons of lame turkeys with swollen joints and tenosynovitis. These TARVs were genetically different from chicken arthritis reoviruses (CARVs) and produced gastrocnemius tenosynovitis when inoculated into turkey poults. The purpose of this study was to determine the pathogenicity of TARVs in chickens. Oneweek-old, specific-pathogen-free chicks were inoculated with either a TARV (TARV-MN2 or TARV-O'Neil) or CARV via oral, intratracheal, or footpad routes. At 2 and 3 weeks post inoculation (PI), a subset of chicks from each group was euthanized followed by collection of tissues for real-time RT-PCR (rRT-PCR), virus isolation, and histopathology. Chickens inoculated with CARV via intratracheal and footpad routes developed gastrocnemius lymphocytic tenosynovitis at 2 and 3 weeks PI. Both TARV-MN2 and TARV-O'Neil induced gastrocnemius lymphocytic tenosynovitis in chicks inoculated only via the footpad route at 2 and 3 weeks PI. Although there was no evidence of clinical lameness, the virus was present in leg tendons, internal organs, and intestines of all TARV-inoculated chicks regardless of route of inoculation, as indicated by rRT-PCR and virus isolation. These results indicate that TARVs do not produce gastrocnemius tenosynovitis in chicks by 3 weeks PI when administered via the most probable natural route (e.g., oral and intratracheal). Further studies are needed to determine the long term effects these viruses might play in inducing lameness in chickens.
Epidemiological survey on Mycoplasma synoviae infection in Portuguese broiler breeder flocks
Since modernization and expansion of the poultry industry, infections with Mycoplasma spp. bacteria have been reported as a cause of considerable economic losses. The prevalence of Mycoplasma synoviae infection in 974,000 Portuguese broiler breeders, belonging to 36 flocks, was investigated from December 2008 to March 2012. This study was conducted using a commercial indirect enzyme-linked immunosorbent assay (ELISA) for the analysis of serum antibodies, and a polymerase chain reaction (PCR) for the tracheal tissue. Twenty-four flocks were simultaneously found positive by ELISA and PCR [66.7%, 95% confidence interval (CI): 43.5-76.9%]. The M. synoviae prevalence among chickens averaged 40.3% (483/1,200), with values ranging from 0.0 to 83.3% per flock. The prevalence of farms where M. synoviae positive birds have been found was determined in different poultry categories such as density, biosecurity, strains, offspring quality, premises'age, and others husbandry factors. Prevalence values were significantly higher among birds housed in new facilities (less than 3 years old) and were also significantly higher in the production period. The high prevalence of M. synoviae infection detected in the present study suggests the need to adopt appropriate control measures.
Iranian Journal of Veterinary Medicine, 2013
BACKGROUND:Avian reoviruses (ARVs) are members of the Orthoreovirus genus; one of the 12 genera of the Reoviridae family. The ARVs are the cause of some important diseases in poultry such as reovirus-induced arthritis, tenosynovitis, chronic respiratory disease, and mal-absorption syndrome. OBJECTIVES: In this study, the presence of ARVs in the Iranian breeder flocks was investigated through reverse transcription- polymerase chain reaction (RT-PCR) and restriction enzyme fragment length polymorphism (RFLP). METHODS: A total of 800 fecal swab samples were initially collected from breeder flocks (older than 45 weeks of age). They were then sent to the laboratory in containers with PBS, and after that they were pooled and finally to 120 samples were obtained. The total RNA extracted from the pooled fecal samples were used to amplify the selected parts of the S1 (1023 bp) and S4 (437 bp) genes from the ARV field isolates using RT-PCR. The positive RT-PCR amplified products were further ...
Seroprevalence of Mycoplasma synoviae infection in broiler breeder and commercial layer fl ocks
Indian Journal of Veterinary Pathology, 47(3): 257-260. , 2023
Seroprevalence Mycoplasma synoviae infection was carried out in male and female breeder fl ocks as well as commercial layer fl ocks by ELISA antibody (Idexx Laboratories USA) testing kit for M. Synoviae. The birds from few healthy fl ocks were also tested for MS antibody. A total of 134 serum samples were collected from ten diff erent male parent fl ocks showing signs of lameness due to tenosynovitis out of which 95 serum samples (70.90%) were positive for MS antibodies. The overall mean titre, titre range and coeffi cient of variation (CV) from male broiler parent (BP) fl ocks were 4123, 0 to 8130 and 70.1, respectively. Moreover, total of 126 serum samples were collected from twelve female BP fl ocks out of which 66 serum samples (52.38%) were positive for MS antibodies. The overall mean titre in female BP fl ocks was 2351 with a CV of 100.4. The titres of these fl ocks were ranged from 11 to 7056. The overall mean titre in two commercial layer fl ocks was 9381 with a titre range of 3693 to 16291 and a CV of 42.3. However, all the serum samples collected from diff erent healthy fl ocks were negative for MS antibodies.
Investigation of Mycoplasma Synoviae Seroprevalence in Broiler Breeder Farms in South Bačka Region
Archives of Veterinary Medicine, 2017
Mycoplasma synoviae is known to cause respiratory disorders, synovitis, subclinical infections, air sacculitis and eggshell apex abnormalitiesin domestic poultry worldwide. Th e aim of this study was to determine M.synoviae seroprevalence in 5 diff erent broiler breeder farms in South Bačka from 2014 to 2017. A total of 1511 samples were tested using commercial indirect enzyme linked immunosorbent assay (ELISA) for detection of antibodies in the blood sera. In this study, the seroprevalence of 25.21% was found and 47 (40.87%) fl ocks out of 115 tested were positive to M.synoviae. Seroprevalence varied between 31.02% in 2015 and 16.78% in 2016. Flock prevalence ranged from 31.03% in 2014 to 55.88% in 2015. Th ese results suggest that M. synoviae infection is present in broiler breeder farms in South Bačka, and that is necessary to conduct further research, systematic monitoring and to improve biosecurity measures on broiler breeder farms.
Test profiles of broiler breeder flocks housed in farms with endemic Mycoplasma synoviae infection
Revista Brasileira de Ciência Avícola, 2003
There is a need for a better understanding of the epidemiology of Mycoplasma synoviae (MS) infection in broiler breeders in Brazil. Many features of the infection remain unrecognizable, because there are no clinical signs of the disease. A detailed testing was performed at each 6 to 8 weeks in three MS-free flocks introduced in farms with endemic MS infection for a follow-up epidemiological study. Every flock was monitored by polymerase chain reaction (PCR), by serum plate agglutination (SPA) and hemagglutination inhibition (HI) for serology studies, and isolation of mycoplasmas from tracheal swabs. PCR was found to be the most sensitive test, detecting early MS infection. Serology was positive in less than 50% of the sera and MS was isolated only between 27 and 28 weeks of age and in a maximum of 60% positive hens. A similar profile was seen for MS infection in all three flocks. Infection started at brooding, whereas laboratory detection of the assymptomatic infection was more probable in the weeks of increasing egg production. This predictable profile during rearing may be very useful for the optimization of monitoring MS infection in broiler breeder flocks.
Detection of Reticuloendotheliosis Virus in Muscovy Ducks, Wild Turkeys, and Chickens in Brazil
Journal of Wildlife Diseases, 2020
Reticuloendotheliosis retroviruses (REVs) are known to cause immunosuppressive and oncogenic disease that affects numerous avian species. Reticuloendotheliosis retroviruses are present worldwide and recently have been reported in South America with cases of infected commercial flocks in Argentina. We surveyed for the presence of REV in birds from a state in the northern region of Brazil using real-time PCR. We report here the presence of REV in Brazil, detected in Muscovy Ducks ( Cairina moschata), Wild Turkeys ( Meleagris gallopavo), and chickens ( Gallus gallus) at a relatively high prevalence (16.8%). Phylogenetic analysis indicated a close relationship of these strains to variants in the United States. This study provides evidence of REV in the Amazon biome and provides a baseline for future surveillance of the virus in the region and throughout Brazil.